Graduate Student Seoul National University, Seoul-t'ukpyolsi, Republic of Korea
Abstract: Pluripotent stem cells (PSCs) are divided into ‘naïve state’ and ‘primed state’ depending on their differentiation potential. Naïve PSCs represent an earlier developmental state with minimal epigenetic features, greater developmental capacity, and enhanced genomic stability than primed PSCs. These features make naïve PSCs more advantageous in studying developmental biology and for applications requiring high-efficiency genome editing. Notably, porcine naïve embryonic stem cells (ESCs) hold significant value due to their physiological similarity to humans and potential use in transgenic animal production. We have recently established porcine primed embryonic stem cells (ESCs); however, there have been no reports on the successful derivation of naïve ESCs in pigs. This is due to the species-specific characteristics of the pluripotency gene network and the variation in the pluripotent state. Therefore, we aimed to identify a species-specific naïve pluripotency gene network in pigs through chemical reprogramming of porcine primed ESCs. The results showed that the reprogrammed cells had dome-shaped colonies, characteristic of naïve PSCs. Interestingly, leukemia inhibitory factor (LIF) and activin a (Act A) appeared to play a key role in porcine naïve pluripotency similar to human naïve PSCs. These cells exhibited alkaline phosphatase activity and the expression of Oct4, Sox2, Nanog, SSEA-1, and SSEA-4. Significant upregulation of naïve PSC markers such as STELLA(P < 0.05) and NANOG(P < 0.001) were also observed compared to porcine primed ESCs. Taken together, we found that porcine primed ESCs can be reprogrammed to the naïve-like state using chemical agents without genetic manipulation. This study will not only be used as preliminary research for establishing embryo-derived porcine naïve ESCs but can also be applied for comparative developmental studies between various species.
Funding Source: This work was supported by the BK21 Four program, the Korea Evaluation Institute of Industrial Technology (KEIT) [20012411]; and the National Research Foundation of Korea (NRF) grant [2023R1A2C1005026].
